Tilbrook A J, Canny B J, Serapiglia M D, Ambrose T J, Clarke I J
Department of Physiology, Monash University, Clayton, Victoria 3168, Australia.
J Endocrinol. 1999 Mar;160(3):469-81. doi: 10.1677/joe.0.1600469.
In this study we used an isolation/restraint stress to test the hypothesis that stress will affect the secretion of LH differently in gonadectomised rams and ewes treated with different combinations of sex steroids. Romney Marsh sheep were gonadectomised two weeks prior to these experiments. In the first experiment male and female sheep were treated with vehicle or different sex steroids for 7 days prior to the application of the isolation/restraint stress. Male sheep received either i.m. oil (control rams) or 6 mg testosterone propionate injections every 12 h. Female sheep were given empty s.c. implants (control ewes), or 2x1 cm s.c. implants containing oestradiol, or an intravaginal controlled internal drug release device containing 0.3 g progesterone, or the combination of oestradiol and progesterone. There were four animals in each group. On the day of application of the isolation/restraint stress, blood samples were collected every 10 min for 16 h for the subsequent measurement of plasma LH and cortisol concentrations. After 8 h the stress was applied for 4 h. Two weeks later, blood samples were collected for a further 16 h from the control rams and ewes, but on this day no stress was imposed. In the second experiment, separate control gonadectomised rams and ewes (n=4/group) were studied for 7 h on 3 consecutive days, when separate treatments were applied. On day 1, the animals received no treatment; on day 2, isolation/restraint stress was applied after 3 h; and on day 3, an i. v. injection of 2 microg/kg ACTH1-24 was given after 3 h. On each day, blood samples were collected every 10 min and the LH response to the i.v. injection of 500 ng GnRH administered after 5 h of sampling was measured. In Experiment 1, the secretion of LH was suppressed during isolation/restraint in all groups but the parameters of LH secretion (LH pulse frequency and amplitude) that were affected varied between groups. In control rams, LH pulse amplitude, and not frequency, was decreased during isolation/restraint whereas in rams treated with testosterone propionate the stressor reduced pulse frequency and not amplitude. In control ewes, isolation/restraint decreased LH pulse frequency but not amplitude. Isolation/restraint reduced both LH pulse frequency and amplitude in ewes treated with oestradiol, LH pulse frequency in ewes treated with progesterone and only LH pulse amplitude in ewes treated with both oestradiol and progesterone. There was no change in LH secretion during the day of no stress. Plasma concentrations of cortisol were higher during isolation/restraint than on the day of no stress. On the day of isolation/restraint maximal concentrations of cortisol were observed during the application of the stressor but there were no differences between groups in the magnitude of this response. In Experiment 2, isolation/restraint reduced the LH response to GnRH in rams but not ewes and ACTH reduced the LH response to GnRH both in rams and ewes. Our results show that the mechanism(s) by which isolation/restraint stress suppresses LH secretion in sheep is influenced by sex steroids. The predominance of particular sex steroids in the circulation may affect the extent to which stress inhibits the secretion of GnRH from the hypothalamus and/or the responsiveness of the pituitary gland to the actions of GnRH. There are also differences between the sexes in the effects of stress on LH secretion that are independent of the sex steroids.
在本研究中,我们采用隔离/束缚应激来验证以下假说:应激对性腺切除的公羊和母羊促黄体生成素(LH)分泌的影响,会因所给予的不同性类固醇组合而有所不同。在这些实验开始前两周,对罗姆尼沼泽羊进行性腺切除。在第一个实验中,在施加隔离/束缚应激前7天,对公羊和母羊分别给予溶剂或不同的性类固醇。公羊接受肌肉注射油剂(对照公羊)或每12小时注射6毫克丙酸睾酮。母羊接受皮下空植入物(对照母羊)、或含有雌二醇的2×1厘米皮下植入物、或含有0.3克孕酮的阴道内可控缓释给药装置、或雌二醇与孕酮的组合。每组有4只动物。在施加隔离/束缚应激当天,每10分钟采集一次血样,持续16小时,用于随后测定血浆LH和皮质醇浓度。8小时后施加4小时应激。两周后,从对照公羊和母羊中再采集16小时血样,但当天不施加应激。在第二个实验中,对单独的性腺切除对照公羊和母羊(每组n = 4)连续3天进行7小时的研究,并施加不同的处理。第1天,动物不接受处理;第2天,3小时后施加隔离/束缚应激;第3天,3小时后静脉注射2微克/千克促肾上腺皮质激素1 - 24。每天每10分钟采集一次血样,并测定在采血5小时后静脉注射500纳克促性腺激素释放激素(GnRH)时的LH反应。在实验1中,所有组在隔离/束缚期间LH分泌均受到抑制,但受影响的LH分泌参数(LH脉冲频率和幅度)在各组之间有所不同。在对照公羊中,隔离/束缚期间LH脉冲幅度而非频率降低,而在用丙酸睾酮处理的公羊中,应激源降低了脉冲频率而非幅度。在对照母羊中,隔离/束缚降低了LH脉冲频率而非幅度。隔离/束缚降低了用雌二醇处理的母羊的LH脉冲频率和幅度、用孕酮处理的母羊的LH脉冲频率以及用雌二醇和孕酮处理的母羊的仅LH脉冲幅度。在无应激日,LH分泌无变化。隔离/束缚期间血浆皮质醇浓度高于无应激日。在施加应激源期间观察到皮质醇的最大浓度,但各组之间该反应的幅度无差异。在实验2中,隔离/束缚降低了公羊对GnRH的LH反应,但未降低母羊的反应,促肾上腺皮质激素降低了公羊和母羊对GnRH的LH反应。我们的结果表明,隔离/束缚应激抑制绵羊LH分泌的机制受性类固醇影响。循环中特定性类固醇的优势可能影响应激抑制下丘脑GnRH分泌的程度和/或垂体对GnRH作用的反应性。应激对LH分泌的影响在性别之间也存在差异,且与性类固醇无关。